Inhibitors of p38 kinase

ABSTRACT

The invention is directed to methods to inhibit p38-α kinase using compounds comprising saturated heterocycles coupled to a fused ring system.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. Ser. No. 09/990,187 filed 20Nov. 2001, and now allowed and to issue as U.S. Pat. No. 6,821,966,which claims benefit under 35 U.S.C. § 119(e) to U.S. Ser. No.60/252,197 filed 20 Nov. 2000. The contents of these applications areincorporated herein by reference.

FIELD OF THE INVENTION

The invention relates to treating various disorders associated withenhanced activity of kinase p38-α. More specifically, it concernscompounds that are related to indole-type derivatives coupled topiperazine- or piperidine-type moieties as useful in these methods.

BACKGROUND ART

A large number of chronic and acute conditions have been recognized tobe associated with perturbation of the inflammatory response. A largenumber of cytokines participate in this response, including IL-1, IL-6,IL-8 and TNF. It appears that the activity of these cytokines in theregulation of inflammation rely at least in part on the activation of anenzyme on the cell signaling pathway, a member of the MAP kinase familygenerally known as p38 and alternatively known as CSBP and RK. Thiskinase is activated by dual phosphorylation after stimulation byphysiochemical stress, treatment with lipopolysaccharides or withproinflammatory cytokines such as IL-1 and TNF. Therefore, inhibitors ofthe kinase activity of p38 are useful anti-inflammatory agents.

Eye diseases associated with a fibroproliferative condition includeretinal reattachment surgery accompanying proliferativevitreoretinopathy, cataract extraction with intraocular lensimplantation, and post glaucoma drainage surgery.

PCT applications WO 98/06715, WO 98/07425, and WO 96/40143, all of whichare incorporated herein by reference, describe the relationship of p38kinase inhibitors with various disease states. As mentioned in theseapplications, inhibitors of p38 kinase are useful in treating a varietyof diseases associated with chronic inflammation. These applicationslist rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, goutyarthritis and other arthritic conditions, sepsis, septic shock,endotoxic shock, Gram-negative sepsis, toxic shock syndrome, asthma,adult respiratory distress syndrome, stroke, reperfusion injury, CNSinjuries such as neural trauma and ischemia, psoriasis, restenosis,cerebral malaria, chronic pulmonary inflammatory disease, silicosis,pulmonary sarcosis, bone resorption diseases such as osteoporosis,graft-versus-host reaction, Crohn's disease, ulcerative colitisincluding inflammatory bowel disease (IBD) and pyresis.

The above-referenced PCT applications disclose compounds which are p38kinase inhibitors said to be useful in treating these disease states.These compounds are either imidazoles or are indoles substituted at the3- or 4-position with a piperazine ring linked through a carboxamidelinkage. Additional compounds which are conjugates of piperazines withindoles are described as insecticides in WO 97/26252, also incorporatedherein by reference.

Certain aroyl/phenyl-substituted piperazines and piperidines whichinhibit p38-α kinase are described in PCT publication WO 00/12074published 9 Mar. 2000. In addition, indolyl substituted piperidines andpiperazines which inhibit this enzyme are described in PCT publicationNo. WO 99/61426 published 2 Dec. 1999. Carbolene derivatives ofpiperidine and piperazine as p38-α inhibitors are described inPCT/US00/07934 filed 24 Mar. 2000.

None of the foregoing patents describes the indole derivatives describedherein which specifically inhibit p38-α.

DISCLOSURE OF THE INVENTION

The invention is directed to methods and compounds useful in treatingconditions that are characterized by enhanced p38-α activity. Theseconditions include inflammation, proliferative diseases, and certaincardiovascular disorders as well as Alzheimer's disease as furtherdescribed below.

Compounds of the invention have been found to inhibit p38 kinase, theα-isoform in particular, and are thus useful in treating diseasesmediated by these activities. The compounds of the invention are of theformula

-   -   and the pharmaceutically acceptable salts thereof, or a        pharmaceutical composition thereof, wherein

represents a single or double bond;

-   -   B is —W_(i)—COX_(j)Y wherein Y is COR² or an isostere thereof        and R² is hydrogen or a noninterfering substituent, each of W        and X is a spacer of 2-6 Å, and each of i and j is independently        0 or 1;    -   each R³ is independently a noninterfering substituent, where n        is 0-3;    -   Z³ is NR⁷ or 0; wherein R⁷ is H or a noninterfering substituent;    -   one Z² is CA or CR⁸A and the other is CR¹, CR¹ ₂, NR⁶ or N        wherein each R¹, R⁶ and R⁸ is independently hydrogen or        noninterfering substituent; wherein A is:

-   -   such that Z¹ is CR⁵ or N wherein R⁵ is hydrogen or a        noninterfering substituent;    -   each of l and k is an integer from 0-2 wherein the sum of 1 and        k is 0-3;    -   Ar is an aryl group substituted with 0-5 noninterfering        substituents, wherein two noninterfering substituents can form a        fused ring;    -   each R⁴ is independently a noninterfering substituent where m is        0-4;    -   each of L¹ and L² is a linker; and    -   the distance between the atom of Ar linked to L² and the center        of the β ring is 4.5-24 Å.

Modes of Carrying out the Invention

The compounds of formula (1) are useful in treating conditions which arecharacterized by overactivity of p38 kinase, in particular theα-isoform. Conditions “characterized by enhanced p38-α activity” includethose where this enzyme is present in increased amount or wherein theenzyme has been modified to increase its inherent activity, or both.Thus, “enhanced activity” refers to any condition wherein theeffectiveness of these proteins is undesirably high, regardless of thecause.

The compounds of the invention are useful in conditions where p38-αkinase shows enhanced activity. These conditions are those in whichfibrosis and organ sclerosis are caused by, or accompanied by,inflammation, oxidation injury, hypoxia, altered temperature orextracellular osmolarity, conditions causing cellular stress, apoptosisor necrosis. These conditions include ischemia-reperfusion injury,congestive heart failure, progressive pulmonary and bronchial fibrosis,hepatitis, arthritis, inflammatory bowel disease, glomerular sclerosis,interstitial renal fibrosis, chronic scarring diseases of the eyes,bladder and reproductive tract, bone marrow dysplasia, chronicinfectious or autoimmune states, spinal chord injury and traumatic orsurgical wounds. These conditions, of course, would be benefited bycompounds which inhibit p38-α. Methods of treatment with the compoundsof the invention are further discussed below.

The Invention Compounds

The compounds useful in the invention are derivatives of indole-typecompounds containing a mandatory substituent, B, preferably at aposition corresponding to the 5 position of the indole nucleus and amandatory substituent A at a position corresponding to the 2- or3-position of indole. In general, an indole-type nucleus is preferred,although alternatives within the scope of the invention are alsoillustrated below.

In the description above, certain positions of the molecule aredescribed as permitting “noninterfering substituents.” This terminologyis used because the substituents in these positions generally speakingare not relevant to the essential activity of the molecule taken as awhole. A wide variety of substituents can be employed in thesepositions, and it is well within ordinary skill to determine whether anyparticular arbitrary substituent is or is not “noninterfering.”

As used herein, a “noninterfering substituent” is a substituent whichleaves the ability of the compound of formula (1) to inhibit p38-αactivity qualitatively intact. Thus, the substituent may alter thedegree of inhibition of p38-α. However, as long as the compound offormula (1) retains the ability to inhibit p38-α activity, thesubstituent will be classified as “noninterfering.” A number of assaysfor determining the ability of any compound to inhibit p38-α activityare available in the art. A whole blood assay for this evaluation isillustrated below: the gene for p38-α has been cloned and the proteincan be prepared recombinantly and its activity assessed, including anassessment of the ability of an arbitrarily chosen compound to interferewith this activity. The essential features of the molecule are tightlydefined. The positions which are occupied by “noninterferingsubstituents” can be substituted by conventional organic moieties as isunderstood in the art. It is irrelevant to the present invention to testthe outer limits of such substitutions. The essential features of thecompounds are those set forth with particularity herein.

In addition, L¹ and L² are described herein as linkers. The nature ofsuch linkers is less important that the distance they impart between theportions of the molecule. Typical linkers include alkylene, i.e.,(CH₂)_(n)—R; alkenylene—i.e., an alkylene moiety which contains a doublebond, including a double bond at one terminus. Other suitable linkersinclude, for example, substituted alkylenes or alkenylenes, carbonylmoieties, and the like.

As used herein, “hydrocarbyl residue” refers to a residue which containsonly carbon and hydrogen. The residue may be aliphatic or aromatic,straight-chain, cyclic, branched, saturated or unsaturated. Thehydrocarbyl residue, when so stated however, may contain heteroatomsover and above the carbon and hydrogen members of the substituentresidue. Thus, when specifically noted as containing such heteroatoms,the hydrocarbyl residue may also contain carbonyl groups, amino groups,hydroxyl groups and the like, or contain heteroatoms within the“backbone” of the hydrocarbyl residue.

As used herein, “inorganic residue” refers to a residue that does notcontain carbon. Examples include, but are not limited to, halo, hydroxy,NO₂ or NH₂.

As used herein, the term “alkyl,” “alkenyl” and “alkynyl” includestraight- and branched-chain and cyclic monovalent substituents.Examples include methyl, ethyl, isobutyl, cyclohexyl, cyclopentylethyl,2-propenyl, 3-butynyl, and the like. Typically, the alkyl, alkenyl andalkynyl substituents contain 1-10C (alkyl) or 2-10C (alkenyl oralkynyl). Preferably they contain 1-6C (alkyl) or 2-6C (alkenyl oralkynyl). Heteroalkyl, heteroalkenyl and heteroalkynyl are similarlydefined but may contain 1-2 O, S or N heteroatoms or combinationsthereof within the backbone residue.

As used herein, “acyl” encompasses the definitions of alkyl, alkenyl,alkynyl and the related hetero-forms which are coupled to an additionalresidue through a carbonyl group.

“Aromatic” moiety refers to a monocyclic or fused bicyclic moiety suchas phenyl or naphthyl; “heteroaromatic” also refers to monocyclic orfused bicyclic ring systems containing one or more heteroatoms selectedfrom O, S and N. The inclusion of a heteroatom permits inclusion of5-membered rings as well as 6-membered rings. Thus, typical aromaticsystems include pyridyl, pyrimidyl, indolyl, benzimidazolyl,benzotriazolyl, isoquinolyl, quinolyl, benzothiazolyl, benzofuranyl,thienyl, furyl, pyrrolyl, thiazolyl, oxazolyl, imidazolyl and the like.Any monocyclic or fused ring bicyclic system which has thecharacteristics of aromaticity in terms of electron distributionthroughout the ring system is included in this definition. Typically,the ring systems contain 5-12 ring member atoms.

Similarly, “arylalkyl” and “heteroalkyl” refer to aromatic andheteroaromatic systems which are coupled to another residue through acarbon chain, including substituted or unsubstituted, saturated orunsaturated, carbon chains, typically of 1-6C. These carbon chains mayalso include a carbonyl group, thus making them able to providesubstituents as an acyl moiety.

When the compounds of Formula 1 contain one or more chiral centers, theinvention includes optically pure forms as well as mixtures ofstereoisomers or enantiomers.

With respect to the portion of the compound between the atom of Ar boundto L² and ring β, L¹ and L² are linkers which space the substituent Arfrom ring a at a distance of 4.5-24 Å, preferably 6-20 Å, morepreferably 7.5-10 Å. The distance is measured from the center of the βring to the atom of Ar to which the linker L² is attached. Typical, butnonlimiting, embodiments of L¹ and L² are CO and isosteres thereof, oroptionally substituted isosteres, or longer chain forms. L², inparticular, may be alkylene or alkenylene optionally substituted withnoninterfering substituents or L¹ or L² may be or may include aheteroatom such as N, S or O. Such substituents include, but are limitedto, a moiety selected from the group consisting of alkyl, alkenyl,alkynyl, aryl, arylalkyl, acyl, aroyl, heteroaryl, heteroalkyl,heteroalkenyl, heteroalkynyl, heteroalkylaryl, NH-aroyl, halo, OR, NR₂,SR, SOR, SO₂R, OCOR, NRCOR, NRCONR₂, NRCOOR, OCONR₂, RCO, COOR,alkyl-OOCR, SO₃R, CONR₂, SO₂NR₂, NRSO₂NR₂, CN, CF₃, R₃Si, and NO₂,wherein each R is independently H, alkyl, alkenyl or aryl or heteroformsthereof, and wherein two substituents on L² can be joined to form anon-aromatic saturated or unsaturated ring that includes 0-3 heteroatomswhich are O, S and/or N and which contains 3 to 8 members or said twosubstituents can be joined to form a carbonyl moiety or an oxime,oximeether, oximeester or ketal of said carbonyl moiety.

Isosteres of CO and CH₂, include SO, SO₂, or CHOH. CO and CH₂ arepreferred.

Thus, L² is substituted with 0-2 substituents. Where appropriate, twooptional substituents on L² can be joined to form a non-aromaticsaturated or unsaturated hydrocarbyl ring that includes 0-3 heteroatomssuch as O, S and/or N and which contains 3 to 8 members. Two optionalsubstituents on L² can be joined to form a carbonyl moiety which can besubsequently converted to an oxime, an oximeether, an oximeester, or aketal.

Ar is aryl, heteroaryl, including 6-5 fused heteroaryl, cycloaliphaticor cycloheteroaliphatic that can be optionally substituted. Ar ispreferably optionally substituted phenyl.

Each substituent on Ar is independently a hydrocarbyl residue (1-20C)containing 0-5 heteroatoms selected from O, S and N, or is an inorganicresidue. Preferred substituents include those selected from the groupconsisting of alkyl, alkenyl, alkynyl, aryl, arylalkyl, acyl, aroyl,heteroaryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heteroalkylaryl,NH-aroyl, halo, OR, NR₂, SR, SOR, SO₂R, OCOR, NRCOR, NRCONR₂, NRCOOR,OCONR₂, RCO, COOR, alkyl-OOCR, SO₃R, CONR₂, SO₂NR₂, NRSO₂NR₂, CN, CF₃,R₃Si, and NO₂, wherein each R is independently H, alkyl, alkenyl or arylor heteroforms thereof, and wherein two of said optional substituents onadjacent positions can be joined to form a fused, optionally substitutedaromatic or nonaromatic, saturated or unsaturated ring which contains3-8 members. More preferred substituents include halo, alkyl (1-4C) andmore preferably, fluoro, chloro and methyl. These substituents mayoccupy all available positions of the aryl ring of Ar, preferably 1-2positions, most preferably one position. These substituents may beoptionally substituted with substituents similar to those listed. Ofcourse some substituents, such as halo, are not further substituted, asknown to one skilled in the art.

Two substituents on Ar can be joined to form a fused, optionallysubstituted aromatic or nonaromatic, saturated or unsaturated ring whichcontains 3-8 members.

Between L¹ and L² is a piperidine-type moiety of the following formula:

Z¹ is CR⁵ or N wherein R⁵ is H or a noninterfering substituent. Each ofl and k is an integer from 0-2 wherein the sum of l and k is 0-3. Thenoninterfering substituents R⁵ include, without limitation, halo, alkyl,alkoxy, aryl, arylalkyl, aryloxy, heteroaryl, acyl, carboxy, or hydroxy.Preferably, R⁵ is H, alkyl, OR, NR₂, SR or halo, where R is H or alkyl.Additionally, R⁵ can be joined with an R⁴ substituent to form anoptionally substituted non-aromatic saturated or unsaturated hydrocarbylring which contains 3-8 members and 0-3 heteroatoms such as O, N and/orS. Preferred embodiments include compounds wherein Z¹ is CH or N, andthose wherein both l and k are 1.

R⁴ represents a noninterfering substituent such as a hydrocarbyl residue(1-20C) containing 0-5 heteroatoms selected from O, S and N. PreferablyR⁴ is alkyl, alkoxy, aryl, arylalkyl, aryloxy, heteroalkyl, heteroaryl,heteroarylalkyl, RCO, ═O, acyl, halo, CN, OR, NRCOR, NR, wherein R is H,alkyl (preferably 1-4C), aryl, or hetero forms thereof. Each appropriatesubstituent is itself unsubstituted or substituted with 1-3substituents. The substituents are preferably independently selectedfrom a group that includes alkyl, alkenyl, alkynyl, aryl, arylalkyl,acyl, aroyl, heteroaryl, heteroalkyl, heteroalkenyl, heteroalkynyl,heteroalkylaryl, NH-aroyl, halo, OR, NR₂, SR, SOR, SO₂R, OCOR, NRCOR,NRCONR₂, NRCOOR, OCONR₂, RCO, COOR, alkyl-OOCR, SO₃R, CONR₂, SO₂NR₂,NRSO₂NR₂, CN, CF₃, R₃Si, and NO₂, wherein each R is independently H,alkyl, alkenyl or aryl or heteroforms thereof and two of R⁴ on adjacentpositions can be joined to form a fused, optionally substituted aromaticor nonaromatic, saturated or unsaturated ring which contains 3-8members, or R⁴ is ═O or an oxime, oximeether, oximeester or ketalthereof. R⁴ may occur m times on the ring; m is an integer of 0-4.Preferred embodiments of R⁴ comprise alkyl (1-4C) especially two alkylsubstituents and carbonyl. Most preferably R⁴ comprises two methylgroups at positions 2 and 5 or 3 and 6 of a piperidinyl or piperazinylring or ═O preferably at the 5-position of the ring. The substitutedforms may be chiral and an isolated enantiomer may be preferred.

R³ also represents a noninterfering substituent. Such substituentsinclude hydrocarbyl residues (1-6C) containing 0-2 heteroatoms selectedfrom O, S and/or N and inorganic residues. n is an integer of 0-3,preferably 0 or 1. Preferably, the substituents represented by R³ areindependently halo, alkyl, heteroalkyl, OCOR, OR, NRCOR, SR, or NR₂,wherein R is H, alkyl, aryl, or heteroforms thereof. More preferably R³substituents are selected from alkyl, alkoxy or halo, and mostpreferably methoxy, methyl, and chloro. Most preferably, n is 0 and theα ring is unsubstituted, except for L¹ or n is 1 and R³ is halo ormethoxy.

In the ring labeled α, Z³ may be NR⁷ or O— i.e., the compounds may berelated to indole or benzofuran. If C³ is NR⁷, preferred embodiments ofR⁷ include H or optionally substituted alkyl, alkenyl, alkynyl, aryl,arylalkyl, acyl, aroyl, heteroaryl, heteroalkyl, heteroalkenyl,heteroalkynyl, heteroalkylaryl, or is SOR, SO₂R, RCO, COOR, alkyl-COR,SO₃R, CONR₂, SO₂NR₂, CN, CF₃, NR₂, OR, alkyl-SR, alkyl-SOR, alkyl-SO₂R,alkyl-OCOR, alkyl-COOR, alkyl-CN, alkyl-CONR₂, or R₃Si, wherein each Ris independently H, alkyl, alkenyl or aryl or heteroforms thereof. Morepreferably, R⁷ is hydrogen or is alkyl (1-4C), preferably methyl or isacyl (1-4C), or is COOR wherein R is H, alkyl, alkenyl of aryl or heteroforms thereof. R⁷ is also preferably a substituted alkyl wherein thepreferred substituents are form ether linkages or contain sulfinic orsulfonic acid moieties. Other preferred substituents include sulfhydrylsubstituted alkyl substituents. Still other preferred substituentsinclude CONR₂ wherein R is defined as above.

It is preferred that the indicated dotted line represents a double bond;however, compounds which contain a saturated a ring are also includedwithin the scope of the invention.

Preferably, the mandatory substituent CA or CR⁸A is in the 3-position;regardless of which position this substituent occupies, the otherposition is CR¹, CR¹ ₂, NR⁶ or N. CR¹ is preferred. Preferredembodiments of R¹ include hydrogen, alkyl, alkenyl, alkynyl, aryl,arylalkyl, acyl, aroyl, heteroaryl, heteroalkyl, heteroalkenyl,heteroalkynyl, heteroalkylaryl, NH-aroyl, halo, OR, NR₂, SR, SOR, SO₂R,OCOR, NRCOR, NRCONR₂, NRCOOR, OCONR₂, RCO, COOR, alkyl-OOCR, SO₃R,CONR₂, SO₂NR₂, NRSO₂NR₂, CN, CF₃, R₃Si, and NO₂, wherein each R isindependently H, alkyl, alkenyl or aryl or heteroforms thereof and twoof R¹ can be joined to form a fused, optionally substituted aromatic ornonaromatic, saturated or unsaturated ring which contains 3-8 members.Most preferably, R¹ is H, alkyl, such as methyl, most preferably, thering labeled a contains a double bond and CR¹ is CH or C-alkyl. Otherpreferable forms of R¹ include H, alkyl, acyl, aryl, arylalkyl,heteroalkyl, heteroaryl, halo, OR, NR₂, SR, NRCOR, alkyl-OOCR, RCO,COOR, and CN, wherein each R is independently H, alkyl, or aryl orheteroforms thereof.

While the position not occupied by CA is preferred to include CR¹, theposition can also be N or NR⁶. While NR⁶ is less preferred (as in thatcase the ring labeled a would be saturated), if NR⁶ is present,preferred embodiments of R⁶ include H, or alkyl, alkenyl, alkynyl, aryl,arylalkyl, acyl, aroyl, heteroaryl, heteroalkyl, heteroalkenyl,heteroalkynyl, heteroalkylaryl, or is SOR, SO₂R, RCO, COOR, alkyl-COR,SO₃R, CONR₂, SO₂NR₂, CN, CF₃, or R₃Si wherein each R is independently H,alkyl, alkenyl or aryl or heteroforms thereof.

Preferably, CR⁸A or CA occupy position 3- and preferably Z² in thatposition is CA. However, if the α ring is saturated and R⁸ is present,preferred embodiments for R⁸ include H, halo, alkyl, alkenyl and thelike. Preferably R⁸ is a relatively small substituent corresponding, forexample, to H or lower alkyl 1-4C.

B is —W_(i)—COX_(j)Y wherein Y is COR² or an isostere thereof and R² isa noninterfering substituent. Each of W and X is a spacer and may be,for example, optionally substituted alkyl, alkenyl, or alkynyl, each ofi and j is 0 or 1. Preferably, W and X are unsubstituted. Preferably, jis 0 so that the two carbonyl groups are adjacent to each other.Preferably, also, i is 0 so that the proximal CO is adjacent the ring.However, compounds wherein the proximal CO is spaced from the ring canreadily be prepared by selective reduction of an initially glyoxalsubstituted β ring. In the most preferred embodiments of the invention,the α/β ring system is an indole containing CA in position 3- andwherein B is bonded to position 5 and is COCR².

The noninterfering substituent represented by R², when R² is other thanH, is a hydrocarbyl residue (1-20C) containing 0-5 heteroatoms selectedfrom O, S and/or N or is an inorganic residue. Preferred are embodimentswherein R² is H, or is straight or branched chain alkyl, alkenyl,alkynyl, aryl, arylalkyl, heteroalkyl, heteroaryl, or heteroarylalkyl,each optionally substituted with halo, alkyl, heteroalkyl, SR, OR, NR₂,OCOR, NRCOR, NRCONR₂, NRSO₂R, NRSO₂NR₂, OCONR₂, CN, COOR, CONR₂, COR, orR₃Si wherein each R is independently H, alkyl, alkenyl or aryl or theheteroatom-containing forms thereof, or wherein R² is OR, NR₂, SR,NRCONR₂, OCONR₂, or NRSO₂NR₂, wherein each R is independently H, alkyl,alkenyl or aryl or the heteroatom-containing forms thereof, and whereintwo R attached to the same atom may form a 3-8 member ring and whereinsaid ring may further be substituted by alkyl, alkenyl, alkynyl, aryl,arylalkyl, heteroalkyl, heteroaryl, heteroarylalkyl, each optionallysubstituted with halo, SR, OR, NR₂, OCOR, NRCOR, NRCONR₂, NRSO₂R,NRSO₂NR₂, OCONR₂, or R₃Si wherein each R is independently H, alkyl,alkenyl or aryl or the heteroatom-containing forms thereof wherein two Rattached to the same atom may form a 3-8 member ring, optionallysubstituted as above defined.

Other preferred embodiments of R² are H, heteroarylalkyl, —NR₂,heteroaryl, —COOR, —NHRNR₂, heteroaryl-COOR, heteroaryloxy, —OR,heteroaryl-NR₂, —NROR and alkyl. Most preferably R² is isopropylpiperazinyl, methyl piperazinyl, dimethylamine, piperazinyl, isobutylcarboxylate, oxycarbonylethyl, morpholinyl, aminoethyldimethylamine,isobutyl carboxylate piperazinyl, oxypiperazinyl, ethylcarboxylatepiperazinyl, methoxy, ethoxy, hydroxy, methyl, amine, aminoethylpyrrolidinyl, aminopropanediol, piperidinyl, pyrrolidinyl-piperidinyl,or methyl piperidinyl.

Isosteres of COR² as represented by Y are defined as follows.

The isosteres have varying lipophilicity and may contribute to enhancedmetabolic stability. Thus, Y, as shown, may be replaced by the isosteresin Table 1.

TABLE 1 Acid Isosteres

Names of Groups Chemical Structures Substitution Groups (SG) tetrazole

n/a 1,2,3-triazole

H; SCH₃; COCH₃; Br;SOCH₃; SO₂CH₃; NO₂; CF₃;CN; COOMe 1,2,4-triazole

H; SCH₃; COCH₃; Br;SOCH₃; SO₂CH₃; NO₂ imidazole

H; SCH₃; COCH₃; Br;SOCH₃; SO₂CH₃; NO₂

Thus, isosteres include tetrazole, 1,2,3-triazole, 1,2,4-triazole andimidazole.

The compounds of formula (1) may be supplied in the form of theirpharmaceutically acceptable acid-addition salts including salts ofinorganic acids such as hydrochloric, sulfuric, hydrobromic, orphosphoric acid or salts of organic acids such as acetic, tartaric,succinic, benzoic, salicylic, and the like. If a carboxyl moiety ispresent on the compound of formula (1), the compound may also besupplied as a salt with a pharmaceutically acceptable cation.

Synthesis of the Invention Compounds

The following Reaction Scheme disclosed in commonly assigned U.S. Ser.No. 09/575,060 is illustrative of the conversion of a 4-benzylpiperidinyl-indole-5-carboxamide to the glyoxalic acid compound andderivatives thereof.

Of course, the 4-benzyl piperidinyl carbonyl of the illustration atposition 5 may be generalized as

-   -   and the glyoxal type substituent at position 3 can be        generalized to W_(i)COXjY. In the present invention the        positions of the glyoxal and the 4-benzylpiperadinyl may be        reversed such that the glyoxal substituent is present at the 5        position of the indole ring and the benzyl pipiperidinyl is        present at the 3 position of the indole.

Similarly, embodiments wherein the indole-type moiety is

-   -   can be used in these schemes. Methods to synthesize the        compounds of the invention are, in general, known in the art.

The following general schemes illustrate such methods.

Substituted amino benzoic acid esters such as I can be treated withreagents such as thiomethylacetaldehyde dimethyl acetal andN-chlorosuccinamide in methylene chloride at low temperature followed bythe treatment with a base such as triethylamine at reflux in methylenechloride, dichloroethane or chloroform to give indoles II, Scheme 1.Treatment with reagents such as Raney-Nickel in an appropriate solventsuch as ethanol, methanol or isopropanol will yield the correspondingindole carboxylic acid ester which when hydrolyzed under base conditionswill give the desired substituted indole carboxylic acid.

Alternatively, substituted amino benzoic acid esters I can be convertedto the ketals IV, Scheme 2, with an appropriate aldehyde underconditions of reductive alkylation with reagents such as sodiumtriacetoxyborohydride in acetic acid in the presence of sodium sulfate.The amines can then be treated with Lewis acids such as aluminumchloride, titanium chloride, BF₃-etherate in dichloromethane ordichloroethane, under reflux to give the corresponding substitutedindole methyl esters, with appropriate substitutions.

Another method could involve the treatment of the substituted aminobenzoic acid esters I with iodine and sodium periodate in an appropriatesolvent such as dimethyformamide, to give the corresponding iodo anilineV, Scheme 3. This can be coupled with an acetylene such as trimethylsilyl acetylene or ethylethynyl ether in the presence of an appropriatecatalysts such as palladium and copper and a base such as triethylamineto give the silyl coupled product such as VI. Subsequent cyclization ina solvent such as dimethylformamide and in the presence of a catalystsuch as copper iodide would give the appropriately substituted indolesVII.

Synthesis of the required piperidines can be achieved by treating anappropriate piperidone such as VIII, Scheme 4, with substituted benzylphosphonate esters in the presence of a base such as sodium hydride togive alkenes which can be reduced to the corresponding substituted4-benzylpiperidine such as IX. The hydrogenations are typically done inthe presence of catalytic metals in solvents such as methanol, ethanoland ethyl acetate.

An alternate method could involve isonipecotoyl chlorides such as Xwhich can be used to acylate appropriately substituted benzenes (ArH) inthe presence of a Lewis acid such as aluminum chloride to give theketones XI, Scheme 5. Further modifications of the carbonyl moiety of XIusing methods and routes generally known can then lead to the desiredcompounds XII.

Substituted piperazines can be reacted with various and appropriateArL²X in the presence or absence of a base or other catalytic reagent togive the substituted piperazines XV, Scheme 6. These can be furtherresolved to the chiral components with the use a chiral resolving agentsuch as tartaric acid to give either enantiomers of the substitutedpiperazines XV.

Compounds III can be treated with halides, acid chlorides and otherelectrophiles (BX), Scheme 7, containing a variety of differentsubstituents, in the presence of a base such as sodium hydride, in avariety of different solvents, to give compounds of type XVI. These canthen be converted to the corresponding acids XVII by treatment withappropriate reagents such as an aqueous base. The acids are then coupledto substituted amines IX, XII or XV using a coupling agent such asEDAC.HCl in a variety of solvents including methylene chloride, dimethylformamide, to give compounds XVIII.

Compounds XVIII can be first treated with acid chlorides such as oxalylchloride in methylene chloride under anhydrous conditions followed bytreatment with a variety of nucleophiles WH to give compounds of typeXIX, Scheme 8.

Compounds of type XXIV can be synthesized starting with theappropriately substituted benzofurans of type XX and coupling them withamines IX, XII or XV in the presence of relevant coupling reagents togive compounds XXI, Scheme 9. Subsequent acylation to XXII can beachieved with an acylating agent such as acetic anhydride in thepresence of a catalyst such as Fe(III). Oxidation of the acetyl moietyof XXII to the glyoxalic acid moiety of XXIII can be accomplished usingan oxidizing agent such as selenium dioxide (Da Settimo, F., et al.,Eur. J. Med. Chem (1996) 31:951-956; Cook, M. C., et al., (1975) Brpatent 1,399,089; Campaigne, E., et al., J. Med. Chem. (1965) 136-137).Finally coupling of the acid with the appropriate nucleophile in WH canbe achieved using any one of the variety of coupling agents known in avariety of solvents to give compounds of type XXIV. Appropriatemodification in the reactions discussed above may be made in accordancewith well know techniques so that the positions of the glyoxal andbenzyl piperadinyl moieties are reversed from that shown above.

Assays for p38 α Kinase Inhibition

For each of the assay procedures described below, the TNF-α productioncorrelates to the activity of p38-α kinase.

A. Human Whole Blood Assay for p38 Kinase Inhibition

Venous blood is collected from healthy male volunteers into aheparinized syringe and is used within 2 hours of collection. Testcompounds are dissolved in 100% DMSO and 1 μl aliquots of drugconcentrations ranging from 0 to 1 mM are dispensed into quadruplicatewells of a 24-well microtiter plate (Nunclon Delta SI, AppliedScientific, So. San Francisco, Calif.). Whole blood is added at a volumeof 1 ml/well and the mixture is incubated for 15 minutes with constantshaking (Titer Plate Shaker, Lab-Line Instruments, Inc., Melrose Park,Ill.) at a humidified atmosphere of 5% CO₂ at 37° C. Whole blood iscultured either undiluted or at a final dilution of 1:10 with RPMI 1640(Gibco 31800+NaHCO₃, Life Technologies, Rockville, Md. and Scios, Inc.,Sunnyvale, Calif.). At the end of the incubation period, 10 μl of LPS(E. coli 0111:B4, Sigma Chemical Co., St. Louis, Mo.) is added to eachwell to a final concentration of 1 or 0.1 μg/ml for undiluted or 1:10diluted whole blood, respectively. The incubation is continued for anadditional 2 hours. The reaction is stopped by placing the microtiterplates in an ice bath and plasma or cell-free supernates are collectedby centrifugation at 3000 rpm for 10 minutes at 4° C. The plasma samplesare stored at −80° C. until assayed for TNF-α levels by ELISA, followingthe directions supplied by Quantikine Human TNF-α assay kit (R&DSystems, Minneapolis, Minn.).

IC₅₀ values are calculated using the concentration of inhibitor thatcauses a 50% decrease as compared to a control.

B. Enriched Mononuclear Cell Assay for p38 Kinase Inhibition

The enriched mononuclear cell assay, the protocol of which is set forthbelow, begins with cryopreserved Human Peripheral Blood MononuclearCells (HPBMC's) (Clonetics Corp.) that are rinsed and resuspended in awarm mixture of cell growth media. The resuspended cells are thencounted and seeded at 1×10⁶ cells/well in a 24-well microtitre plate.The plates are then placed in an incubator for an hour to allow thecells to settle in each well.

After the cells have settled, the media is aspirated and new mediacontaining 100 ng/ml of the cytokine stimulatory factorLipopolysaccharide (LPS) and a test chemical compound is added to eachwell of the microtiter plate. Thus, each well contains HPBMC's, LPS anda test chemical compound. The cells are then incubated for 2 hours, andthe amount of the cytokine Tumor Necrosis Factor Alpha (TNF-α) ismeasured using an Enzyme Linked Immunoassay (ELISA). One such ELISA fordetecting the levels of TNF-α is commercially available from R&DSystems. The amount of TNF-α production by the HPBMC's in each well isthen compared to a control well to determine whether the chemicalcompound acts as an inhibitor of cytokine production.

LPS Induced Cytokine Synthesis in HPBMC's

-   -   Cryopreserved HPBMC (cat#CC-2702 Clonetics Corp)    -   LGM-3 media (cat#CC-3212 Clonetics Corp)    -   LPS stock 10 μg/ml (Cat. No. L 2630 serotype 0111:B4 Sigma)    -   Human TNF-α ELISA (R&D Systems)    -   DNase I (10 mg/ml stock)

Preparation of Cells.

-   -   LGM-3 media warmed to 37° C.    -   5 μl of DNase I stock added to 10 ml media.    -   Cells thawed rapidly and dispersed into above.    -   Centrifuge 200×g×10 min @ RT.    -   Pellet up in 10 ml sterile PBS.    -   Centrifuge 200×g×10 min @ RT.    -   Pellet resuspended in 10 ml LGM-3 then diluted to 50 ml with        LGM-3.    -   Perform cell count.    -   Adjust to 1×E06 cells/well.    -   Seed 1 ml/well of a 24 well plate.    -   Place plate in incubator to plate down for 1 hour.

Preparation of Incubation Media.

-   -   LGM-3 containing 100 ng/ml LPS (e.g., 50 ml media plus 0.5 ml        LPS stock)    -   Aliquot into 2 ml aliquots and add 1000× inhibitor dilutions.

Incubation

When cells have plated down aspirate media away and overlay with 1 mlrelevant incubation media. Return plate to incubator for 2 hours or 24hours. Remove supernatants after incubation to a labeled tube and eitherperform TNF (or other) ELISA immediately or freeze for later assay.

IC₅₀ values are calculated using the concentration of inhibitor thatcauses a 50% decrease as compared to a control.

Administration and Use

The compounds of the invention are useful among other indications intreating conditions associated with inflammation. Thus, the compounds offormula (1) or their pharmaceutically acceptable salts are used in themanufacture of a medicament for prophylactic or therapeutic treatment ofmammals, including humans, in respect of conditions characterized byexcessive production of cytokines and/or inappropriate or unregulatedcytokine activity on such cells as cardiomyocytes, cardiofibroblasts andmacrophages.

The compounds of the invention inhibit the production of cytokines suchas TNF, IL-1, IL-6 and IL-8, cytokines that are importantproinflammatory constituents in many different disease states andsyndromes. Thus, inhibition of these cytokines has benefit incontrolling and mitigating many diseases. The compounds of the inventionare shown herein to inhibit a member of the MAP kinase family variouslycalled p38 MAPK (or p38), CSBP, or SAPK-2. The activation of thisprotein has been shown to accompany exacerbation of the diseases inresponse to stress caused, for example, by treatment withlipopolysaccharides or cytokines such as TNF and IL-1. Inhibition of p38activity, therefore, is predictive of the ability of a medicament toprovide a beneficial effect in treating diseases such as Alzheimer's,coronary artery disease, congestive heart failure, cardiomyopathy,myocarditis, vasculitis, restenosis, such as occurs following coronaryangioplasty, atherosclerosis, IBD, rheumatoid arthritis, rheumatoidspondylitis, osteoarthritis, gouty arthritis and other arthriticconditions, multiple sclerosis, acute respiratory distress syndrome(ARDS), asthma, chronic obstructive pulmonary disease (COPD), silicosis,pulmonary sarcosis, sepsis, septic shock, endotoxic shock, Gram-negativesepsis, toxic shock syndrome, heart and brain failure (stroke) that arecharacterized by ischemia and reperfusion injury, surgical procedures,such as transplantation procedures and graft rejections, cardiopulmonarybypass, coronary artery bypass graft, CNS injuries, including open andclosed head trauma, inflammatory eye conditions such as conjunctivitisand uveitis, acute renal failure, glomerulonephritis, inflammatory boweldiseases, such as Crohn's disease or ulcerative colitis, graft vs. hostdisease, bone resorption diseases like osteoporosis, type II diabetes,pyresis, psoriasis, cachexia, viral diseases such as those caused byHIV, CMV, and Herpes, and cerebral malaria.

Within the last several years, p38 has been shown to comprise a group ofMAP kinases designated p38-α, p38-β, p38-γ and p38-δ. Jiang, Y., et al.,J Biol Chem (1996) 271:17920-17926 reported characterization of p38-β asa 372-amino acid protein closely related to p38-α. In comparing theactivity of p38-α with that of p38-β, the authors state that while bothare activated by proinflammatory cytokines and environmental stress,p38-β was preferentially activated by MAP kinase kinase-6 (MKK6) andpreferentially activated transcription factor 2, thus suggesting thatseparate mechanisms for action may be associated with these forms.

Kumar, S., et al., Biochem Biophys Res Comm (1997) 235:533-538 andStein, B., et al., J Biol Chem (1997) 272:19509-19517 reported a secondisoform of p38-β, p38-β2, containing 364 amino acids with 73% identityto p38-α. All of these reports show evidence that p38-β is activated byproinflammatory cytokines and environmental stress, although the secondreported p38-β isoform, p38-β2, appears to be preferentially expressedin the CNS, heart and skeletal muscle compared to the more ubiquitoustissue expression of p38-α. Furthermore, activated transcriptionfactor-2 (ATF-2) was observed to be a better substrate for p38-β2 thanfor p38-α, thus suggesting that separate mechanisms of action may beassociated with these forms. The physiological role of p38-β1 has beencalled into question by the latter two reports since it cannot be foundin human tissue and does not exhibit appreciable kinase activity withthe substrates of p38-α.

The identification of p38-γ was reported by Li, Z., et al., BiochemBiophys Res Comm (1996) 228:334-340 and of p38-δ by Wang, X., et al., JBiol Chem (1997) 272:23668-23674 and by Kumar, S., et al., BiochemBiophys Res Comm (1997) 235:533-538. The data suggest that these two p38isoforms (γ and δ) represent a unique subset of the MAPK family based ontheir tissue expression patterns, substrate utilization, response todirect and indirect stimuli, and susceptibility to kinase inhibitors.

Various results with regard to response to drugs targeting the p38family as between p38-α and either the putative p38-β1 or p38-β2 or bothwere reported by Jiang, Kumar, and Stein cited above as well as byEyers, P. A., et al., Chem and Biol (1995) 5:321-328. An additionalpaper by Wang, Y., et al., J Biol Chem (1998) 273:2161-2168 suggests thesignificance of such differential effects. As pointed out by Wang, anumber of stimuli, such as myocardial infarction, hypertension, valvulardiseases, viral myocarditis, and dilated cardiomyopathy lead to anincrease in cardiac workload and elevated mechanical stress oncardiomyocytes. These are said to lead to an adaptive hypertrophicresponse which, if not controlled, has decidedly negative consequences.Wang cites previous studies which have shown that in ischemiareperfusion treated hearts, p38 MAPK activities are elevated inassociation with hypertrophy and programmed cell death. Wang shows inthe cited paper that activation of p38-β activity results inhypertrophy, whereas activation of p38-α activity leads to myocyteapoptosis. Thus, selective inhibition of p38-α activity as compared top38-β activity will be of benefit in treating conditions associated withcardiac failure. These conditions include congestive heart failure,cardiomyopathy, myocarditis, vasculitis, vascular restenosis, valvulardisease, conditions associated with cardiopulmonary bypass, coronaryartery bypass, grafts and vascular grafts. Further, to the extent thatthe α-isoform is toxic in other muscle cell types, α-selectiveinhibitors would be useful for conditions associated with cachexiaattributed to TNF or other conditions such as cancer, infection, orautoimmune disease.

Thus, the invention encompasses the use of compounds which selectivelyinhibit the activity of the p38-α isoform for treating conditionsassociated with activation of p38-α, in particular those associated withcardiac hypertrophy, ischemia or other environmental stress such asoxidation injury, hyperosmolarity or other agents or factors thatactivate p38-α kinase, or cardiac failure, for example, congestive heartfailure, cardiomyopathy and myocarditis.

The manner of administration and formulation of the compounds useful inthe invention and their related compounds will depend on the nature ofthe condition, the severity of the condition, the particular subject tobe treated, and the judgment of the practitioner; formulation willdepend on mode of administration. As the compounds of the invention aresmall molecules, they are conveniently administered by oraladministration by compounding them with suitable pharmaceuticalexcipients so as to provide tablets, capsules, syrups, and the like.Suitable formulations for oral administration may also include minorcomponents such as buffers, flavoring agents and the like. Typically,the amount of active ingredient in the formulations will be in the rangeof 5%-95% of the total formulation, but wide variation is permitteddepending on the carrier. Suitable carriers include sucrose, pectin,magnesium stearate, lactose, peanut oil, olive oil, water, and the like.

The compounds useful in the invention may also be administered throughsuppositories or other transmucosal vehicles. Typically, suchformulations will include excipients that facilitate the passage of thecompound through the mucosa such as pharmaceutically acceptabledetergents.

The compounds may also be administered topically, for topical conditionssuch as psoriasis, or in formulation intended to penetrate the skin.These include lotions, creams, ointments and the like which can beformulated by known methods.

The compounds may also be administered by injection, includingintravenous, intramuscular, subcutaneous or intraperitoneal injection.Typical formulations for such use are liquid formulations in isotonicvehicles such as Hank's solution or Ringer's solution.

Alternative formulations include nasal sprays, liposomal formulations,slow-release formulations, and the like, as are known in the art.

Any suitable formulation may be used. A compendium of art-knownformulations is found in Remington's Pharmaceutical Sciences, latestedition, Mack Publishing Company, Easton, Pa. Reference to this manualis routine in the art.

The dosages of the compounds of the invention will depend on a number offactors which will vary from patient to patient. However, it is believedthat generally, the daily oral dosage will utilize 0.001-100 mg/kg totalbody weight, preferably from 0.01-50 mg/kg and more preferably about0.01 mg/kg-10 mg/kg. The dose regimen will vary, however, depending onthe conditions being treated and the judgment of the practitioner.

It should be noted that the compounds of formula (1) can be administeredas individual active ingredients, or as mixtures of several embodimentsof this formula. In addition, the inhibitors of p38 kinase can be usedas single therapeutic agents or in combination with other therapeuticagents. Drugs that could be usefully combined with these compoundsinclude natural or synthetic corticosteroids, particularly prednisoneand its derivatives, monoclonal antibodies targeting cells of the immunesystem, antibodies or soluble receptors or receptor fusion proteinstargeting immune or non-immune cytokines, and small molecule inhibitorsof cell division, protein synthesis, or mRNA transcription ortranslation, or inhibitors of immune cell differentiation or activation.

As implied above, although the compounds of the invention may be used inhumans, they are also available for veterinary use in treating animalsubjects.

The following examples are intended to illustrate but not to limit theinvention, and to illustrate the use of the above Reaction Schemes.

EXAMPLE 1

Preparation of B. To a solution of 4-nitrophenylglyoxalic acid (2.0 g,10.25 mmol) in 30 mL CH₂Cl₂ was added 5.2 mL of 2.0 molar solution ofN,N-dimethylamine in THF (10.4 mmol) and1-[3-dimethylamino)propyl]3-ethylcarbodiimide (2.0 g, 10.25 mmol). Themixture was stirred at rt for 4 h. TLC showed the reaction was complete.The solvent was evaporated and the residue was dissolved in EtOAc. Theorganic layer was washed with water, and brine. Dried with Na₂SO₄ andevaporated to get 2.1 g (93%) of the product as yellow solids. MS 222(M⁺).

Preparation of C. To a solution of B (1.8 g, 8.1 mmol) in 10 mL of EtOHwas added Sn(II)Cl₂.2H₂O (9.1 g, 40.5 mmol) followed by 5 mL of con.HCl. The mixture was heated to 60° C. for 3 h whereby the startingmaterial was completely disappeared as seen by TLC and MS. Evaporated toa residue with was carefully basified with 3N NaOH and extracted withEtOAc. The organic layer was dried with Na₂SO₄ and evaporated to get1.08 g (70%) of the product as yellow solids. MS 192 (M⁺).

Preparation of D. A mixture of compound C (1.0 g, 5.2 mmol), anhydroussodium sulfate (7.4 g, 52.0 mmol), and pyruvic aldehyde dimethyl acetal(1.3 g, 10.4 mmol) in 40 ml of AcOH was stirred at rt for 10 min. Thensodium triacetoxyborohydride (3.3 g, 15.6 mmol) was added in portionsfor 5 min and stirred for 2 h. The reaction was complete as seen by TLCand MS. The reaction mixture was filtered and the filtrate wasconcentrated at reduced pressure. The residue was dissolved in EtOAc andwashed several times with saturated sodium carbonate. The organic layerwas dried with Na₂SO₄ and evaporated. The crude product was purified bysilica gel chromatography using EtOAc:hexane (1:1) as eluent. CompoundsD (1.2 g, 80%) was obtained as white solids. MS 294 (M⁺).

Preparation of E. To a suspension of AlCl₃ (2.1 g, 16.0 mmol) in 20 mLof dichloroethane was added a solution of compound D (1.1 g, 4.0 mmol)in 10 mL dichloroethane and stirred at rt for 1 h, then warmed to 60° C.for an additional 1 h. All starting material was disappeared accordingto TLC. Cooled, quenched with methanol, evaporated, and extracted withEtOAc. The organic layer was dried with Na₂SO₄ and evaporated. The crudeproduct was purified by silica gel chromatography using EtOAc:hexane(1:1) as eluent. Compound E (0.37 g, 45%) was obtained as white solids.MS 230 (M⁺).

Preparation of Compound F. To a solution of indole E (0.1 g, 0.44 mmol)in 5 ml of CH₂Cl₂ was added pyridine (0.44 mmol) and the mixture wascooled to 0° C. Then a tolune solution of phosgene (1.0 mmol) was addedand stirred at 0° C. for 1 h and then at rt overnight. Evaporated andtreated with a solution of 4-fluorobenzylpiperidine (1.0 mmol) inCH₂Cl₂. The product was purified by silica gel chromatography usingCH₂Cl₂MeOH to obtain 100 mg (50%) of product as white solids, MS 449(M⁺).

EXAMPLE 2

Compound G was prepared using the same method as shown for compound F inExample 1 above.

EXAMPLE 3 Synthesis of(2S,5R)N-4-Fluorobenzyl-trans-2,5-dimethylpiperazine

(+/−)N-Benzyl-trans-2,5-dimethylpiperazine was synthesized as follows

50 g trans-2,5 dimethylpiperazine was dissolved in 300 mL ethanol andtreated with 26.36 mL (½ equivalents) benzylbromide. The mixture wasstirred at room temperature for 12 hours and concentrated. The residuewas taken up in ethyl acetate and washed with 10% aqueous sodiumbicarbonate and saturated sodium chloride; dried over anhydrousmagnesium sulfate and concentrated to give crude1-benzyl-trans-2,5-dimethylpiperazine, as an oil.

This material was chromatographed using DCM/MeOH 95/5 to remove thedi-alkylated product and then with DCM/MeOH/TEA 90/10/0.1 to elute thebenzyl-trans-2,5-dimethylpiperazine. 19.6 g of the pure product wasobtained as an oil.

To a solution of (+/−)N-benzyl-trans-2,5-dimethylpiperazine (59 g, 0.29mol) in methanol (150 mL) was added a solution of (+) tartaric acid (87g, 0.58 mol) in methanol (250 mL) dropwise over 5 min. Crystallizationis effected by keeping the resulting mixture at 0° C. for 48-72 hours.Scratching of the solution after 12-16 hours facilitates thecrystallization process. The mixture was filtered and washed with coldmethanol and dried to give the ditartaric acid salt (73.9 g) as whitecrystals. A single recrystallization from methanol, cooling to roomtemperature afforded the salt as white crystals. (58 g) [α]_(D)=+47,(c=1.00, methanol).

31 g, 15.27 mmol, dimethylbenzylpiperazine was treated with 43 g, 19.85mmol of di-tert-butyl-dicarbonate in 250 mL THF for 4 hours. Thereaction was monitored by TLC and is essentially complete when theaddition of (BOC)₂O is complete. The solvent was removed and the residuewas taken up in ethyl acetate and washed with 10% aqueous sodiumcarbonate and saturated sodium chloride, dried over anhydrous sodiumsulfate and concentrated to give 39.3 g of the Boc-protected compound.This material was used for the next step without further purification.

39.3 g, 131 mmol, of the Boc-protected benzylpiperazine was treated with3.93 g of Pearlman's catalyst, in 150 mL methanol with 3 mL acetic acidfor 4 hours at 40 psi, hydrogen pressure in a parr shaker. The reactionmixture was filtered through celite and concentrated to give a residuethat was dried under high vacuum and then dissolved in 250 mL dryethanol and treated with 1.2 equivalents, 158 mmol of4-Fluorobenzylbromide, and 2 equivalents, 262 mmol of triethylamine for5 hours. The reaction mixture was monitored by TLC and found to becomplete at that time. The solvent was removed and the residue was takenup in ethyl acetate and washed with 10% aqueous sodium carbonate andsaturated sodium chloride, dried over anhydrous sodium sulfate andconcentrated to give 47 g of crude product. This crude material waschromatographed on silica gel using Hexane/Ethyl acetate 95/5 to elutethe product. After chromatography 29 g of the4-Fluorobenzyl-trams-2,5-dimethylpiperazine was obtained.

20 g of the 1-(4-Fluoro)benzyl-trams-2,5-dimethylpiperazine, 62 mmol wastreated with a mixture 200 mL 4 N HCl/Dioxane/2 M HCl/Ether (1:3) for 1hour. Electron impact mass spectroscopy confirmed the formation ofproduct and the disappearance of starting material at that time. Thereaction mixture was concentrated to give a white solid. This materialwas repeatedly washed with hexane and ether to remove residual dioxane,and then dried under vacuum extensively before use. 17.6 g of the finalproduct was obtained as a white fluffy solid.

ADDITIONAL EXAMPLES

Synthesis of 2: Methyl 4-amino-2-chlorobenzoate (1) (18.5 g) wasdissolved in dichloromethane (350 ml) and methyl thioacetaldehydedimethylacetal (13.6 g) was added. The mixture was cooled to −45° C.(dry ice/acetonitrile bath). N-chlorosuccinimide (16.0 g) in 350 mldichloromethane was added dropwise over 1 hr 30 min while maintainingbath temp at −45° C. The reaction mixture was stirred additional 1 hr,then triethylamine (16 mL, 100 mmols) in 30 ml dichloromethane was addeddropwise over 5 min, reaction was warmed to room temp, then refluxed for16 h. Solvent was removed and residue taken up in 500 ml carbontetrachloride, triethylamine-hydrochloric acid was removed byfiltration, filtrate was heated to reflux for 2 h. Solvent was removedby rotary evaporation.

The residue was dissolved in 250 ml tetrahydrofuran and 250 ml 10%hydrochloric acid was added. The mixture was stirred overnight at roomtemperature until the complete disappearance of the starting materialwas observed. Solvent was removed under vacuum, acidic aqueous solutionwas extracted with ethyl acetate (3×125 ml). The combined ethyl acetateextracts were washed with 10% hydrochloric acid, water and dried overanhydrous sodium sulfate. Solvent was removed under vacuum. Crudeproduct mixture was purified on a silica column eluting with ethylacetate:hexanes (15:85) to give 6.4 g of the desired product 2.

Synthesis of 3: Methyl 6-Chloro-3-thiomethyl-5-indole carboxylate (5.2g) was dissolved in 150 ml ethanol:tetrahydrofuran (9:3) and treatedwith Raney-Nickel. Reaction was monitored by mass spec at 30 minintervals, with subsequent addition of Raney-Nickel until reaction wascomplete. When reaction was complete reaction was carefully filteredthrough celite and the celite washed with methanol several times andfiltrate evaporated. Residue was taken up in ethyl acetate, washed withwater, dried over anhydrous sodium sulfate. The solvent was removed togive 3 (3.2 g).

Synthesis of 4: Methyl ester 1.5 g was dissolved in 30 ml methanol/water50:50. The reaction mixture was heated at 50° C. for 2 h with 4 Mol.equivalent sodium hydroxide. The reaction mixture was cooled inice-bath, acidified to pH 3 with 5M hydrochloric acid, removed methanolby rotary evaporation and extracted with ethyl acetate. The extract waswashed with saturated sodium chloride and dried over anhydrous sodiumsulfate. Evaporation of the solvent gave the desired acid 4 (1.48 g).

Synthesis of 5: STEPA: The phosphonate A (38.4 g) and the piperidone B(35.4) were dissolved in anhydrous dimethylformamide (400 mL). To thissodium hydride (60% suspension in oil) was added in portions while thereaction is maintained at 0° C. After the addition of sodium hydride wascomplete the reaction mixture was stirred for 30 min. and then the icebath was removed, the reaction was allowed to stir for 6 h as it slowlywarmed to ambient temperature. The reaction was again cooled in an icebath and quenched with methanol. Water was added to the reactionmixture, and the product extracted with ethyl acetate. The ethyl acetatelayer was washed with saturated sodium chloride and dried over anhydrousmagnesium sulfate. The solvent was removed to gives the crude alkene,which is purified by column chromatography eluting with ethylacetate/hexane (1:9) to give 21.8 g of the desired product C.

Synthesis of 5: STEP B: 10.1 g of C was dissolved in 50 mL methanol.After purging the solution with nitrogen, 5% Palladium on carbon (1 g)catalyst was added followed by 1 mL acetic acid. The parr containercontaining the reaction mixture was hydrogenated for 4 h at 40-50 psi.The reaction mixture was filtered through celite and concentrated. Theresidue was treated with 2 M hydrochloric acid in ether to convert tothe hydrochloric acid salt. The white solid that was obtained was driedunder vacuum, extensively, to give 7.8 g of 5 as the hydrochloric acidsalt.

Synthesis of 6: A mixture of 6-chloro-indole-5-carboxylic acid (1.95 g),4-fluoro-benzylpiperidine hydrochloric acid salt (2.76 g) was taken in50 mL dry dichloromethane and was treated with triethylamine (1.7 mL).The mixture was stirred until a clear solution was obtained.1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (2.3 g) anddimethylaminopyridine (0.25 g) was added and the mixture was stirred for20 h at ambient temperature. The mixture was poured in to water and theorganic layer separated. The aq. Solution was further extracted twicewith dichloromethane. The combined extract was washed with 10%hydrochloric acid. The organic layer was dried over magnesium sulfateand evaporated. The product was purified by chromatography on silica geleluting with ethylacetate:hexane (3:7) to give 6 (2.78 g).

Synthesis of 7: 8.61 g of6-chloro-(4-F-benzylpiperidinyl)-indole-5-carboxamide (6) was dissolvedin about 100 mL dry DMF and the solution was cooled in an ice-bath. Tothe cold solution was added 30 mL of 1M solution of sodiumbis-(trimethylsilyl)-amide in tetrahydrofuran under inert atmosphere.The reaction was allowed to stir at 0° C. for 15 min and at ambienttemperature for another 30 min. The reaction mixture was cooled again inice-bath and 2.5 mL of iodomethane was added. After stirring for 30 min.at 0° C., it was allowed to warm to AMBIENT TEMPERATURE and stirringcontinued for 18 h at AMBIENT TEMPERATURE. The mixture was diluted withwater (or brine) and the product was extracted with ethylacetate (4×75mL). The combined extract was washed with water and dried over anh.MgSO₄. The solvent was removed and the product was purified by columnchromatography on silica gel eluting with ethylacetate:hexane (1:4) toyield 8.0 g of the desired product 7.

Synthesis of 8: 8 g of 7 was dissolved in about 100 mL of anhydrousdichloromethane and was cooled in an ice-bath. To this a 2M solution ofoxalylchloride (20.8 mL) in dichloromethane was added slowly via asyringe and the mixture was allowed to stir at 0° C. for 1 h. Theice-bath was removed and stirring continued for an additional 2 h atAMBIENT TEMPERATURE. The solvent was removed under reduced pressure andthe residue was pumped for 15 min. to remove any excess oxalylchloridepresent. The product was immediately redissolved in anh. dichloromethane(150 mL), cooled in an ice-bath, 30 mL of a 2M solution of dimethylaminein tetrahydrofuran was added rapidly via a syringe. After 15 min.stirring it was allowed to stir for another 15 min. at ambienttemperature. The solution was washed with water to remove the salt, anddried. After evaporation, the residue was purified on silica gel,eluting with chloroform:methanol (99:1) to yield 9.3 g of 8.

Synthesis of 10: STEP A: To a solution of dimethyl piperazine 9 (25 g)in 300 ml of absolute ethanol was added 400 ml of 2N hydrogen chloridein diethyl ether. The solution was warmed to 70° C. in an oil bath for20 minutes. The solution was then cooled to room temperature and set at6° C. overnight. The solid obtained, was collected by filtration. Yield39.8 g (dihydrochloride salt of trans-2,5 dimethylpiperazine) afterdrying overnight under high vacuum.

Synthesis of 10: STEP B: An ethanol solution of 42.9 g of dimethylpiperazine dihydrochloride the from STEP A and 26.1 g trans-2,5dimethylpiperazine was vigorously stirred in an oil bath at 80° C. untilall starting materials were dissolved. The temperature of oil bath wasreduced to 65° C. and 33.1 g of 4-fluro benzylchloride was added. Afterstirring at this temperature for 30 min., the solution was placed in a6° C. refrigerator overnight. The solid was removed from the solution byfiltration and excess of 2N hydrogen chloride in diethyl ether was addedto the filtrate. The filtrate was kept at 6° C. overnight and the solidcollected. The solid was suspended in 5% sodium hydroxide aqueoussolution and extracted three times with ethyl acetate. The organic layerwas dried over sodium sulfate and dried down to give a yellow oil.

Synthesis of 10: STEP C: A solution of 50.7 g (L)-tartaric acid in 130ml of boiling methanol was added to 70 ml of hot methanol solution of37.5 g of the product from STEP B. The solution was set at 6° C. for 96hours before collection of white fine crystals by filtration.

This material was recrystallized from boiling methanol. The product wascollected by filtration after being kept at a 6° C. overnight. Yield30.5 g of ditartaric acid salt ([α]=+43.2°, c=1).

Synthesis of 11: The indole ester 3 (0.526 g) was dissolved in 10 mLacetone (dry) and placed in an ice bath. To this was added crushedpotassium hydroxide (0.7 g, 12.5 mmol), after stirring for five minutesat 0° C., methyl iodide (400 μL, 6.272 mmol) was added to the reactionmixture. The reaction mixture was stirred at 0° C. for 10 minutes andthen at room temperature for 30 minutes. After removal of the solvent,the residue was taken up in ethyl acetate and washed with saturatedsodium chloride. After drying over anhydrous sodium sulfate, filtrationand rotary evaporation, a solid was obtained, 0.7 g. Columnchromatography on silica with ethyl acetate:hexane (2:8) gave methylester of 11 as a white solid. 0.52 g. 0.52 g of this product wasdissolved in 50 mL methanol and treated with 5 mL 10 N sodium hydroxide,the reaction mixture was heated at 50 for 2 h. The reaction mixture wascooled to room temperature and concentrated to a solid on a rotaryevaporator. The residue was taken in 50 mL water, washed with ether andplaced in an ice bath. The basic solution was acidified with 10%hydrochloric acid to pH 2. The precipitate was extracted with ethylacetate and the ethyl acetate layer was washed with saturated sodiumchloride solution. Drying over anhydrous sodium sulfate, filtration andconcentration on a rotary evaporator to gave 11 (0.48 g) as a whitesolid.

Synthesis of 12: 1.56 g of the acid (11) was dissolved in dry methylenechloride 10 mL and to this was added the1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (1.57 g) anddimethylaminopyridine (catalytic). After stirring under nitrogen for 10min., 2.19 g of the amine (10) was added, followed by triethylamine (3mL). After stirring overnight at room temperature, the reaction mixturewas concentrated and the residue was taken up in ethyl acetate andwashed with 10% aq. sodium carbonate, aq. saturated sodium chloride,dried over anhydrous sodium sulfate and filtered. Concentration gave thecrude product which was chromatographed on silica gel using ethylacetate:hexane (4:6) as eluent to give 1.02 g of the desired product.

Synthesis of 13: 1.02 g of 12 from the previous step was dissolved in 30mL dry dichloromethane. The reaction mixture was purged with nitrogenand placed in an ice bath. To this was added 4 mL of 2M oxalyl chloridein dichloromethane. The reaction mixture was stirred at 0° C. for 1 hourand then at room temperature for 2 h. Reaction mixture was concentratedon a rotary evaporator. After drying on a vacuum pump for 15 min. theresidue (a yellow solid) was dissolved in dry dichloromethane (30 mL) towhich was added 4 mL of a 2M solution of dimethylamine intetrahydrofuran. 30 minutes later the reaction mixture was concentratedand the residue was taken up in ethyl acetate and washed with 10% aq.sodium carbonate, saturated sodium chloride, dried over anhydrous sodiumsulfate and filtered. Concentration gives the crude product that waschromatographed on silica gel with ethyl acetate:methanol (9:1). Thewhite solid obtained was dissolved in 10 mL dry dichloromethane. To thiswas added sufficient 2 M hydrochloric acid in ether, till a precipitatepersisted. The mixture was then concentrated on a rotary evaporator todryness and then further dried overnight under high vacuum to give 13(1.08 g).

Synthesis of 15: To a solution of aniline 1 (9.25 g, 0.05 mol) andpyruvic aldehyde dimethyl acetal 14 (11.8 g, 0.1 mol) in 200 mL glacialacetic acid was added anhydrous sodium sulfate (71.0 g, 0.5 mol) and themixture was stirred for 30 min. Powdered sodium triacetoxy borohydride(31.8 g, 0.15 mol) was then added in portions for a period of 5 min. Thereaction mixture was stirred for an additional 2 h. Acetic acid wasremoved under reduced pressure and the residue was made basic by addingsufficient amount of saturated sodium bicarbonate solution. The productwas then extracted with ethyl acetate, dried with sodium sulfate andevaporated to get an oil. This was chromatographed on silica gel columnusing ethyl acetate:hexane (3:7) to give 15 (14 g) as colorless oil.

Synthesis of 16 and 17: To a suspension of fresh aluminum chloride (18.5g) in 200 ml dry chloroform at 0° C. was added a solution of ketal 15(13.3 g) in 100 ml chloroform slowly and the mixture was allowed to warmup to the room temperature and stirred overnight. Ice-cold water wasadded carefully to quench the aluminum chloride and the organic layerwas separated and washed with sodium bicarbonate solution, dried andevaporated to get a white solid. The isomers were separated using silicagel column chromatography using ethyl acetate:hexane (1:9). The 6-choloindole 17 (2.0 g) eluted first followed by 4-chloro isomer 16 (3.8 g).

Synthesis of 18. To a solution of 1.3 g of indole 17 in 15 mL ofmethanol was added a solution of 0.9 g of sodium hydroxide in 20 mL ofwater. The reaction mixture was heated at 50° C. for 4 h where upon aclear solution resulted. Cooled and evaporated off methanol and theresidue was diluted with water and acidified with 10% hydrochloric acid.The product was extracted with ethyl acetate. The organic layer wasdried over sodium sulfate, filtered and evaporated to obtain indole acid18 (1.2 g) as white solid.

2-Methyl-6-methoxyindole-5-carboxylic acid was also synthesized usingthe above synthetic procedure.

Synthesis of 21: To a solution of methyl 4-amino-6-methoxy 5-benzoate(19) (6.0 g, 0.033 mol) and dimethyl acetal 20 (7.0 g, 0.066 mol) in 150mL glacial acetic acid was added anhydrous sodium sulfate (47.0 g, 0.33mol) and the mixture was stirred for 30 min. Powdered sodium triacetoxyborohydride (20.1 g, 0.099 mol) was then added in portions for a periodof 5 min. The reaction mixture was stirred for an additional 2 h. Aceticacid was removed under reduced pressure and the residue was made basicby adding sufficient amount of saturated sodium bicarbonate solution.The product was then extracted with ethyl acetate, washed with saturatedsodium chloride, dried over sodium sulfate and evaporated to get an oil.This was chromatographed on silica gel column using ethyl acetate:hexane(3:7) as eluent and the desired product 21 was obtained (5.2 g) as anoil.

Synthesis of 22: To a solution of 21 (3.6 g) and iodomethane (5.7 g) in50 mL anhydrous dimethylforamide was added potassium t-butoxide (1.0 Min tetrahydrofuran, 20 mL) at ambient temperature. The reaction mixturewas stirred at ambient temperature for 0.5 h and poured into 250 mLethyl acetate, washed with water (4×100 mL), brine (50 mL) and driedover magnesium sulfate. Evaporation of solvent afforded 3.26 g of 22.The product was used for next step without purification.

Synthesis of 23: To a suspension of anhydrous aluminum chloride (0.71 g)in 20 mL anhydrous 1,2-dichloroethane was added, dropwise a solution of22 (1 g) in 10 mL 1,2-dichloroethane with stirring. The reaction washeated to 80° C. for 0.5 h. At the end of this time, the reactionmixture was quenched with methanol, solvents evaporated, then ethylacetate (100 mL) was added. The organic phase was washed with water, aq.sodium bicarbonate and brine and concentrated. The crude product waspurified by silica chromatography using ethyl acetate:hexane (3:7) togive 23 0.22 g.

Synthesis of 24. To a suspension of 1.2 g of indole acid 18 and 1.6 g of5 in 30 mL dichloromethane was added 0.7 g of triethylamine followed by1.4 g of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide. The clearsolution obtained was stirred for 4 h. The solvent was evaporated andthe residue was taken up in ethyl acetate and washed with water, dil.hydrochloric acid, and brine. The organic layer was then dried withsodium sulfate and evaporated. The product was isolated (1.3 g) as whitesolids after silica gel chromatography using ethyl acetate:hexane (1:4).

Synthesis of 25: A solution of 1.3 g of 24 in 20 mL dichloromethane wascooled to 0° C. and a solution of 2 M oxalyl chloride in dichloromethane(3.4 ml, 6.8 mmol) was added and the reaction mixture was stirred at 0°C. for 1 h. The temperature was allowed to come to room temperature andcontinued stirring for an additional 1 h. The solvent was evaporated andthe residue was dried under vacuum. The acid chloride was dissolved indichloromethane (25 mL) and a solution of 6.8 mL (13.6 mmol) of 2 MN,N-dimethylamine in tetrahydrofuran was added all at once. The solventwas removed and the product was purified by silica gel columnchromatography using methanol:chloroform (2:98). The product wasobtained as white solid (1.4 g).

Synthesis of 27: A solution of 26 (130 mg) in anhydrous dimethyl for (5ml) was stirred at 0° C. under nitrogen. Sodium hydride (25 mg, 60%dispersion in oil) was added and stirred for 5 min., then room temp for30 min. The mixture was cooled at 0° C. and chloromethyl methyl sulfide(70 μl) was added. The reaction mixture was stirred for 20 h at roomtemp, and then was extracted with ethyl acetate. The organic layer wasdried over sodium sulfate, concentrated under vacuum. The residue waspurified by chromatotron using methanol:chloroform (2:98) to give 100 mgof 27.

Synthesis of 28: A solution of oxone (120 mg) in water (1 ml) was addedto the suspension of 27 (100 mg) in glacial acetic acid (4 ml). Thereaction mixture was stirred at room temp. for 18 h, washed with water,and concentrated under vacuum. The residue was purified by chromatotronusing methanol:chloroform (2:98) to give 60 mg of 28.

Synthesis of 30: To a solution of 29 (200 mg) in tetrahydrofuran (20 mL)was added sodium bis(trimethylsilyl)amide (0.51 mL, 0.51 mmol) dropwiseat 0° C. The mixture became light yellow solution, warmed to ambienttemperature slowly and was stirred for 30 min. Then methoxy methylchloride was added and the reaction mixture was stirred overnight. Thereaction mixture was quenched with ammonium chloride and extracted withdichloromethane. The combined organic layer was washed with brine, driedand concentrated. The residue was purified by chromatography on silicagel and eluted with methanol:dichloromethane (5:95) to give 190 mg of30.

Synthesis of 31: To a solution of 29 (200 mg) in tetrahydrofuran (10 mL)was added sodium bis(trimethylsilyl)amide (0.56 mL, 0.56 mmol) dropwiseat 0° C. The reaction mixture was warmed to ambient temperature slowly,stirred for 30 min, and then added to the solution of tosyl cyanide (115mg) in tetrahydrofuran (10 mL). The resulting mixture was stirred for 2h at rt, quenched with ammonium chloride and extracted withdichloromethane. The combined organic layer was washed with brine, driedand concentrated. The residue was purified by chromatography on silicagel and eluted with methanol:dichloromethane (5:95) to give 90 mg of 31.

Synthesis of 34: Piperidone 33 was added to a solution of 4-flurobenzylmagnesium chloride (33) (33 mL, 8.3 mmol, 0.25 M in ethyl ether) slowlyat 0° C. The reaction mixture was warmed up to ambient temperature andthen at reflux for 6 h. The resulting milky solution was treated withammonium chloride (saturated) and extracted with ether. The combinedorganic layer was washed with brine, dried and concentrated. The residuewas purified by chromatography on silica gel eluting with hexane:ethylacetate (4:1) to give 990 mg of 34.

Synthesis of 35: The alcohol 34 (670 mg) in dichloromethane (10 mL) wasadded to the solution of diethylaminosulfur trifluride (0.57 mL, 4.34mmol) in dichloromethane (20 mL) at −78° C. The reaction mixture waswarmed up to ambient temperature slowly and stirred for 2 h, and thentreated with sodium carbonate (saturated) and extracted withdichloromethane. The combined organic layer was washed with brine, driedand concentrated. The residue was purified by chromatography on silicagel eluting with hexane:ethyl acetate (8:1) to give 300 mg of 35.

Synthesis of 36: A mixture of the 35 (456 mg) and 4 M of hydrochloricacid in dioxane (10 mL) was stirred for 4 h. The reaction mixture wasneutralized with sodium carbonate, extracted with ethyl acetate. Thecombined organic layer was washed with brine, dried and concentrated.The crude product was used in the next reaction without furtherpurification.

Synthesis of 38: To the suspension of indole carboxlic acid 37 (475 mg)in anhydrous dichloromethane (20 mL) was added piperidine 36 (350 mg,1.66 mmol). The mixture was stirred for 10 min and then1-[3-(dimethylaminopropyl]3-ethylcarbodiimide hydrochloride (475 mg) anddimethylaminopyridine (202 mg) were added. The reaction mixture becameclear and was continually stirred for overnight. Then the reactionmixture was treated with 10% of hydrochloric acid solution, andextracted with dichloromethane. The combined organic extracts werewashed with sodium bicarbonate, brine, dried and concentrated. Theresidue was purified by chromatography on silica gel eluting withdichloromethane:ethyl acetate (9:1) to give 300 mg of 38 as a whitefoam.

Synthesis of 39: To the suspension of 38 (200 mg) was added oxalychloride (0.52 mL, 1.04 mmol, 2 M in dichloromethane) dropwise at 0° C.The reaction mixture was stirred at 0° C. for 30 min, and then warmed upto rt. and stirred for 5 h. The yellow suspension was formed. Thesolvent and excess oxaly chloride were removed under the reducedpressure. The yellow solid formed was dried under vacuum, and dissolvedin dichloromethane then cooled in ice bath. Dimethyl amine (1.04 mL,2.08 mmol, 2 M in tetrahydrofuran) was added. After 30 min, the reactionmixture was treated with water and extracted with dichloromethane. Theorganic extracts were washed with water, brine, dried and concentrated.The residue was purified by chromatography on silica gel eluting with 3%of MeOH in dichloromethane to give 190 mg of 39 as a white solid.

Synthesis of 40: To a solution of 39 (75 mg) in tetrahydrofuran (10 mL)was added KHMDS (0.46 mL, 0.23 mmol, 0.5 M in toluene) dropwise at 0° C.The mixture was warmed to rt. slowly and stirred for 30 min. Then methyliodide (33 mg, 0.23 mmol) was added slowly to the reaction mixture, thenstirred for 2 h. The reaction mixture was quenched with ammoniumchloride and extracted with dichloromethane. The combined organicextracts were washed with brine, dried and concentrated. The residue waspurified by chromatography on silica gel eluting withmethanol:dichloromethane (2:98) to give 50 mg of 40.

Synthesis of 41: To a solution of 40 (75 mg) in ^(tetrahydrofuran) (10mL) was added KHMDS (0.46 mL, 0.23 mmol, 0.5 M in toluene) dropwise at0° C. The mixture was warmed to rt. slowly and stirred for 30 min. Thenmethoxy methyl chloride (18 mg, 0.23 mmol) was added slowly to thereaction mixture, then stirred overnight. The reaction mixture wasquenched with ammonium chloride and extracted with dichloromethane. Thecombined organic extracts were washed with brine, dried andconcentrated. The residue was purified by chromatography on silica geleluting with methanol:dichloromethane (2:98) to give 65 mg of 41.

Synthesis of 43: To a suspension of 6-methoxy-5-indolecarboxylic acid(37) (200 mg) in dichloromethane (5 mL) was added1-[3-(dimethylaminopropyl]3-ethylcarbodiimide hydrochloride (258 mg,1.35 mmol) in one portion. The reaction mixture was stirred till all ofthe solid was dissolved. 4-Benzyl-4-hydroxypiperidine (42) (258 mg),obtained by removal of the protecting group from 34 as outlined in thesynthesis of 36, was added and stirred at ambient temperature. Thereaction mixture became cloudy. A catalytic amount ofdimethylaminopyridine (10 mg) was added and stirred at ambienttemperature overnight. The reaction mixture was treated with water, andextracted with dichloromethane. The combined organic layer was washedwith 10% of hydrochloric acid solution, sodium bicarbonate (saturated)and brine, then dried and concentrated to give 300 mg (82%) of 43 as ayellow foam.

Synthesis of 44: Oxalyl chloride (0.41 mL, 1.65 mmol, 2 M indichloromethane) was added to a solution of6-methoxy-(4-benzyl-4-hydroxypiperidinyl)-indole-5-carboxamide (300 mg)in dichloromethane (20 mL) at 0° C. dropwise. The reaction mixture wasstirred at 0° C. for 30 min, and then warmed up to ambient temperatureslowly. After 2 h., a yellow precipitation was formed. The reactionmixture was concentrated under the reduced pressure and dried for 1 hunder reduced pressure. The resulting yellow solid was suspended indichloromethane (20 mL). Methyl piperazine (0.2 mL) and diisopropylethyl amine (0.2 mL) were added at rt. The mixture was stirred for 1 h,treated with water, and extracted with dichloromethane. The residue waspurified by chromatography on silica gel eluting withdichloromethane:methanol (10:1) to give 180 mg of a white solid. 20 mgof this white solid was dissolved in methanol (1 mL). To this asaturated hydrochloric acid solution in methanol was added dropwise tillthe pH value was around 3. Then the solvents were removed and theproduct was dried to give 44.

Synthesis of 45: To a solution of 44 (65 mg) in anhydroustetrahydrofuran (10 mL) was added sodium bis(trimethylsilyl)amide (0.25mL, 0.25 mmol, 1.0 M solution in tetrahydrofuran) dropwise at ambienttemperature. The mixture was stirred at ambient temperature for 30 min,and then ethyl chloroformate (0.024 mL, 0.25 mmol) was added to thereaction mixture. After 1 h, the reaction mixture was quenched withammonium chloride (saturated), and extracted with ethyl acetate. Thecombined organic layer was washed with brine, dried (magnesium sulfate)and concentrated under reduced pressure. The residue was purified bychromatography on silica gel, eluting with dichloromethane:methanol(10:1) to give 15 mg of the title compound 45 as a white solid.

This was dissolved in methanol (1 mL) and to this was added saturatedhydrochloric acid solution in methanol dropwise till the pH value isaround 3. Then the solvent was removed and the product was dried to givethe hydrochloric acid salt of 45.

Synthesis of 47: STEP A: Piperizinone 46 (5.0 g) was dissolved in 15 mLtetrahydrofuran, to this was added a solution of di-tertbuty-dicarbonate(12.04 g in 50 mL tetrahydrofuran). After stirring at room temperaturefor 4 h. The reaction mixture was concentrated to dryness on a rotaryevaporator and the solid obtained used for the next reaction withoutfurther purification.

Synthesis of 47: STEP B: 10 g of product from STEP A was dissolved inanhydrous acetone and the reaction mixture was cooled in an ice bath. Tothis was added crushed potassium hydroxide (14.02 g). After stirring at0° C. for 10 min. 4-fluorobenzylbromide (23.5 g) was added and thereaction mixture stirred at 0° C. for 20 min and at room temperature for1 h. After removal of the solvent on a rotary evaporator the residue wastaken up in ethyl acetate and washed with 10% aqueous sodium carbonate,brine and dried over anhydrous sodium sulfate. Filtration andconcentration gave crude material that was purified by chromatography onsilica gel using ethyl acetate/hexanes gave 14 g of 47.

Synthesis of 48: 14 g of 47 was dissolved in 100 mL methylene chlorideand treated with 100 mL 2 M Hydrochloric acid in ether for 2 h. Thesolvent was removed and the solid obtained was washed with ether andhexanes and dried under high vacuum to give 48 (11.3 g) as thehydrochloride salt.

Synthesis of 49: 1 g of 48, was dissolved in 20 mL methylene chlorideand treated with 6-methoxy-5-indole carboxylic acid (37) (0.87 g),1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (0.95 g), triethylamine(700 μL) and dimethyaminopyridine (catalytic) and the reaction mixturestirred overnight. The reaction mixture was concentrated and the residuetaken up in ethyl acetate, washed with 10% aqueous sodium carbonate,brine and dried over anhydrous sodium sulfate. Filtration andconcentration gave crude material that was purified by chromatography onsilica gel using ethyl acetate/hexanes to give 49 (1.6 g).

Synthesis of 50: 1.6 g of 49 was dissolved in 25 mL dry dichloromethane.The reaction mixture was purged with nitrogen and placed in an ice bath.To this was added, 5 mL 2M oxalyl chloride in dichloromethane. Thereaction mixture was stirred at room temperature for 1 h and then atroom temperature for 2 h. Reaction mixture was concentrated on a rotaryevaporator and after drying on a vacuum pump for 15 min. the residue (ayellow solid) was dissolved in dry dichloromethane. To this residue a 2Msolution of dimethylamine in THF (5 mL) was added and the reactionmixture stirred for 0.5 h. The reaction mixture was then concentratedand the residue was taken up in ethyl acetate and washed with 10% aq.sodium carbonate, saturated sodium chloride, dried over anhydrous sodiumsulfate and filtered. Concentration gives the crude product that waschromatographed on silica gel using ethyl acetate/hexanes to give 50(1.45 g).

Synthesis of 52: 2R,5S-transdimethyl piperazine 51 (5.0 g) was dissolvedin 15 mL ethanol, to this was added 2-bromoethylbenzene (4.4 g). Thereaction mixture was stirred at 40° C. for 30 min., cooled to roomtemperature and concentrated to dryness on a rotary evaporator. Theresidue was taken up in ethyl acetate and washed with 10% aqueous sodiumcarbonate, brine and dried over anhydrous sodium sulfate. Filtration andconcentration gave crude material that was purified by chromatography onsilica gel using ethyl acetate/hexanes gave 52 (5.4 g).

Synthesis of 53: 5 g of 52 was dissolved in 60 mL methylene chloride andtreated with 60 mL 2 M hydrochloric acid in ether for 3 h. The solventwas removed and the solid obtained was washed with ether and hexanes anddried under high vacuum to give 53 as the hydrochloride salt, 3.6 g.

Synthesis of 54: 53 (1.32 g), was dissolved in 20 mL methylene chlorideand treated with 6-methoxy-5-indole carboxylic acid (37) (0.9 g) in thepresence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (1.04 g), 800μL triethylamine and catalytic DMAP, overnight. The reaction mixture wasconcentrated and the residue taken up in ethyl acetate, washed with 10%aqueous sodium carbonate, brine and dried over anhydrous sodium sulfate.Filtration and concentration gave crude material that was purified bychromatography on silica gel using ethyl acetate/hexanes to give 54(0.95 g).

Synthesis of 55: 0.6 g of 54 was dissolved in 20 mL dry methylenechloride. The reaction mixture was purged with nitrogen and placed in anice bath. To this was added, 2 mL of a 2M oxalyl chloride in methylenechloride. The reaction mixture was stirred at room temperature for 1 hand then at room temperature for 2 h. Reaction mixture was concentratedon a rotary and after drying on a vacuum pump for 15 min. the residue (ayellow solid) was dissolved in 15 mL dry methylene chloride, to whichwas added 2 mL of a 2M solution of dimethylamine in tetrahydrofuran. 30minutes later the reaction mixture was concentrated and the residue wastaken up in ethyl acetate and washed with 10% aq. sodium carbonate,saturated sodium chloride, dried over anhydrous sodium sulfate andfiltered. Concentration gives the crude product that was chromatographedon silica gel using ethyl acetate/hexanes to give 55 (0.78 g).

Synthesis of 57: 7.65 g (19.97 mmol) of the indole 56 was dissolved in80 mL anhydrous dimethylformamide and was cooled in an ice-bath. Underan inert atmosphere, 40 mL (2M solution in dichloromethane) ofoxalylchloride was added dropwise over 20 min. After stirring for 15min. at 0° C., the reaction was allowed to warm up to ambienttemperature and stirring continued for 1 h. The reaction mixture waspoured in to water and extracted with ethylacetate (3×100 mL). Thecombined extract was washed with water and dried over sodium sulfate andevaporated. The product was further purified by column chromatographywith ethylacetate:hexane (30:70) to yield 8 g (97.5%) of the aldehyde57.

Synthesis of 58: To a solution of 1-methylimidazole (149 mg) intetrahydrofuran (10 mL) was added n-butyl lithium (1.14 mL, 1.82 mmol,1.6 M in hexane) dropwise at −78° C. The reaction mixture was stirred at−40° C. for 30 min. 57 (500 mg) in tetrahydrofuran (10 mL) was added.The reaction mixture was warmed to ambient temperature and stirredovernight. The reaction mixture was quenched with ammonium chloride(saturated), extracted with ethyl acetate. The combined organic layerwas washed with brine, dried and concentrated. The crude mixture wasreflux with phenyl hydrazine (0.6 mL) in ethanol (10 mL) to remove 57,which was not consumed in the reaction. Removal of the solvent, theresidue was purified by chromatography on silica gel eluting with ethylacetate:hexane (4:1) to give 120 mg of 58.

Using the foregoing procedures, and other chemical synthetic methodswell known in the art, the following compounds may be prepared inaccordance with the present invention. All of the compounds areinhibitors of p38α.

1. A compound of the formula:

or the pharmaceutically acceptable salts thereof, wherein

represents a single or double bond; B is—W₁—COX_(j)Y wherein Y istetrazole; 1,2,3-triazole; 1,2,4-triazole; or imidazole or is COR²wherein R² is H, straight or branched chain alkyl, alkenyl, alkynyl,aryl, arylalkyl, heteroalkyl, heteroaryl, or heteroarylalkyl, eachoptionally substituted with halo, alkyl, heteroalkyl, SR, OR, NR₂, OCOR,NRCOR, NRCONR₂, NRSO₂R, NRSO₂NR₂, OCONR₂, CN, COOR, CONR₂, COR, or R₃Siwherein each R is independently H, alkyl, heteroalkyl, alkenyl,heteroalkenyl, aryl or heteroaryl, or wherein R² OR, NR₂, SR, NRCONR₂,OCONR₂, or NRSO₂NR₂, wherein each R is independently H, alkyl,heteroalkyl, alkenyl, heteroalkenyl, aryl or heteroaryl, and wherein twoR attached to the same atom may form a 3-8 member ring and wherein saidring may further be substituted by alkyl, alkenyl, alkynyl, aryl,arylalkyl, heteroalkyl, heteroaryl, heteroarylalkyl, each optionallysubstituted with halo, SR, OR, NR₂, OCOR, NRCOR, NRCONR₂, NRSO₂R,NRSO₂NR₂, OCONR₂, or R₃Si wherein each R is independently H, alkyl,heteroalkyl, alkenyl, heteroalkenyl, aryl or heteroaryl wherein two Rattached to the same atom may form a 3-8 member ring, optionallysubstituted as above defined; each of W and X is an alkylene oralkenylene of 2-6Å length, and each of i and j is independently 0 or 1;each R³ is independently halo, alkyl, heteroalkyl, OCOR, OR, NRCOR, SR,or NR₂, wherein R is H, alkyl, heteroalkyl, alkenyl, heteroalkenyl, arylor heteroaryl; n is 0-3; Z³ is NR⁷; wherein R⁷ is H, alkyl, alkenyl,alkynyl, aryl, arylalkyl, acyl, aroyl, heteroaryl, heteroalkyl,heteroalkenyl, heteroalkynyl, heteroalkylaryl, each optionallysubstituted with halo, SR, OR, NR₂, OCOR, NRCOR, NRCONR₂, NRSO₂R,NRSO₂NR₂, OCONR₂, CN, COOR, CONR₂, COR, or R₃Si wherein each R isindependently H, alkyl, heteroalkyl, alkenyl, heteroalkenyl, aryl orheteroaryl and wherein any aryl or heteroaryl group may be substitutedby alkyl or heteroalkyl or is SOR, SO₂R, RCO, COOR, alkyl-COR, SO₃R,CONR₂, SO₂NR₂, CN, CF₃, NR₂, OR, alkyl-SR, alkyl-SOR, alkyl-SO₂R,alkyl-OCOR, alkyl-COOR, alkyl-CN, alkyl-CONR₂, or R₃Si, wherein each Ris independently H, alkyl, heteroalkyl, alkenyl, heteroalkenyl, aryl orheteroaryl; one Z² CA or CR⁸ A and the other is CR¹ or CR¹ ₂, whereineach R¹ is independently hydrogen, alkyl, alkenyl, alkynyl, aryl,arylalkyl, acyl, aroyl, heteroaryl, heteroalkyl, heteroalkenyl,heteroalkynyl, heteroalkylaryl, NH-aroyl, halo, OR, NR₂, SR, SOR, SO₂R,OCOR, NRCOR, NRCONR₂, NRCOOR, OCONR₂, RCO, COOR, alkyl-OOCR, SO₃R,CONR₂, SO₂NR₂, NRSO₂NR₂, CN, CF₃, R₃Si, and NO₂, wherein each R isindependently H, alkyl, heteroalkyl, alkenyl, heteroalkenyl, aryl orheteroaryl and two of R¹ can be joined to form a fused, optionallysubstituted aromatic or nonaromatic, saturated or unsaturated ring whichcontains 3-8 members; R⁸ is H, halo, alkyl or alkenyl; A is:

wherein R⁵ is H, OR, NR₂, SR or halo, wherein each R is independently H,alkyl, heteroalkyl, alkenyl, heteroalkenyl, aryl or heteroaryl; Ar is anaryl group substituted with 0-5 substituents selected from the groupconsisting of alkyl, alkenyl, alkynyl, aryl, arylalkyl, acyl, aroyl,heteroaryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heteroalkylaryl,NH-aroyl, halo, OR, NR₂, SR, SOR, SO₂R, OCOR, NRCOR, NRCONR₂, NRCOOR,OCONR₂, RCO, COOR, alkyl-OOCR, SO₃R, CONR₂, SO₂NR₂, NRSO₂NR₂, CN, CF₃,R₃Si, and NO₂, wherein each R is independently H, alkyl, heteroalkyl,alkenyl, heteroalkenyl, aryl or heteroaryl, and wherein two of saidoptional substituents on adjacent positions can be joined to form afused, optionally substituted aromatic or nonaromatic, saturated orunsaturated ring which contains 3-8 members; each R⁴ is independentlyalkyl, alkenyl, alkynyl, aryl, arylalkyl, acyl, aroyl, heteroaryl,heteroalkyl, heteroalkenyl, heteroalkynyl, heteroalkylaryl, NH-aroyl,halo, OR, NR₂, SR, SOR, SO₂R, OCOR, NRCOR, NRCONR₂, NRCOOR, OCONR₂, RCO,COOR, alkyl-OOCR, SO₃R, CONR₂, SO₂NR₂, NRSO₂NR₂, CN, CF₃, R₃Si, and NO₂,wherein each R is independently H, alkyl, heteroalkyl, alkenyl,heteroalkenyl, aryl or heteroaryl and two of R⁴ on adjacent positionscan be joined to form a fused, optionally substituted aromatic ornonaromatic, saturated or unsaturated ring which contains 3-8 members,or R⁴ is =O or an oxime, oximeether, oximeester or ketal thereof; m is0-4; L¹ is CO, CHOH or CH₂; and L² is alkylene (1-4C) or alkenylene(2-4C) optionally substituted with a moiety selected from the groupconsisting of alkyl, alkenyl, alkynyl, aryl, arylalkyl, acyl, aroyl,heteroaryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heteroalkylaryl,NH-aroyl, halo, OR, NR², SR, SOR, SO₂R, OCOR, NRCOR, NRCONR₂, NRCOOR,OCONR₂, RCO, COOR, alkyl-OOCR, SO₃R, CONR₂, SO₂NR₂, NRSO₂NR₂, CN, CF₃,R₃Si, and NO₂, wherein each R is independently H, alkyl, heteroalkyl,alkenyl, heteroalkenyl, aryl or heteroaryl, and wherein two substituentson L² can be joined to form a non-aromatic saturated or unsaturated ringthat includes 0-3 heteroatoms which are O, S and/or N and which contains3 to 8 members or said two substituents can be joined to form a carbonylmoiety or an oxime, oximeether, oximeester or ketal of said carbonylmoiety.
 2. The compound of claim 1 wherein each of i and j is
 0. 3. Thecompound of claim 1 wherein R⁷ is H, alkyl or acyl each optionallysubstituted with halo, SR, OR, NR₂, OCOR, NRCOR, NRCONR₂, NRSO₂R,NRSO₂NR₂, OCONR₂, CN, COOR, CONR₂, COR, or R₃Si wherein each R isindependently H, alkyl, heteroalkyl, alkenyl, heteroalkenyl, aryl orheteroaryl.
 4. The compound of claim 1 wherein L¹ is CO.
 5. The compoundof claim 1 wherein L² is unsubstituted alkylene.
 6. The compound ofclaim 1 wherein L² is unsubstituted methylene, methylene substitutedwith alkyl, or—CH═.
 7. The compound of claim 1 wherein Ar is optionallysubstituted phenyl.
 8. The compound of claim 7 wherein said optionalsubstitution is by halo, OR, or alkyl.
 9. The compound of claim 8wherein said phenyl is unsubstituted or has a single substituent. 10.The compound of claim 1 wherein each R⁴ is halo, OR, or alkyl.
 11. Thecompound of claim 10 wherein m is 0, 1, or
 2. 12. The compound of claim11 wherein m is 2 and both R⁴ are alkyl.
 13. The compound of claim 1wherein R³ is halo or alkoxy.
 14. The compound of claim 13 wherein n is0, 1 or
 2. 15. The compound of claim 1 wherein L¹ is coupled to the βring at the 5-position.
 16. The compound of claim 1 wherein Z² atposition 3 is CA or CR⁸ A.
 17. The compound of claim 16 wherein the Z²at position 2 is CR¹ or CR¹ ₂.
 18. The compound of claim 17 wherein eachR¹ is selected from the group consisting of H, alkyl, acyl, aryl,arylalkyl, heteroalkyl, heteroaryl, halo, OR, NR₂, SR, NRCOR,alkyl-OOCR, RCO, COOR, and CN, wherein each R is independently H, alkyl,heteroalkyl, alkenyl, heteroalkenyl, aryl or heteroaryl.
 19. Thecompound of claim 1 wherein

represents a double bond.
 20. The compound of claim 1 wherein thecompound of formula (1) is selected from the group consisting of:


21. A pharmaceutical composition which comprises a therapeuticallyeffective amount of a compound of claim 1 in admixture with apharmaceutically acceptable excipient.
 22. A method to treat a conditionselected from the group consisting of multiple sclerosis, inflammatorybowel disease, rheumatoid arthritis, rheumatoid spondylitis,osteoarthritis, gouty arthritis, sepsis, septic shock, endotoxic shock,Gram-negative sepsis, toxic shock syndrome, asthma, adult respiratorydistress syndrome, reperfusion injury, psoriasis, restenosis, cerebralmalaria, chronic pulmonary inflammatory disease, silicosis, pulmonarysarcosis, a bone resorption disease, graft-versus-host reaction, Crohn'sdisease, ulcerative colitis, and pyresis. which method comprisesadministering to a subject in need of such treatment a compound of claim1 or a pharmaceutical composition thereof.